Methods of treating opportunistic infections with azaspiranes

ABSTRACT

Invented is a method of treating opportunistic infections in a mammal, including a human, in need thereof which comprises administering to such mammal an effective amount of a substituted azaspirane.

METHODS

This invention relates to a method of treating opportunistic infectionsin a mammal, including a human, in need thereof which comprisesadministering to such mammal an effective amount of a substitutedazaspirane.

BACKGROUND OF THE INVENTION

Opportunistic infections are an increasing problem in medicine.Opportunistic infections can be caused by a wide variety of bacteria,viruses, fungi and protozoa, as described in Microbiology 16th edition.Appleton Crofts, N.Y., 1976, p 405. Of particular interest are e.g.Candida sp., Pseudomonas sp., Listeria sp., Pneumocystis carinii,Pneumococci, Neisseria sp., Salmonella sp., Mycobacteria sp.,Cryptococcus sp., Aspergillis sp., Cryptosporidium sp., Herpes simplex,Herpes zoster, Cytomegalovirus and Toxoplasma sp. These organisms, whichare often part of the normal flora, are rarely a cause for concern innormal hosts but, under certain circumstances, can cause seriousdisease. These circumstances include but are not limited to: prolongedhigh dose antibiotic therapy, cancer chemotherapy, transplantation andacquired immunodeficiency syndrome (AIDS). In the first two examplescited, a Biologic Response Modifier, (i.e., compounds withimmunostimulatory activity, e.g. muramylpeptides) would be the adjuncttreatment of choice for an opportunistic infection (Walsh TH et al. CurrOpin Oncol 4:647-655, 1992). In the latter two examples, however,immunostimulation may be contraindicated. In the case oftransplantation, a Biologic Response Modifier would be expected toexacerbate graft rejection of graft versus host disease. In AIDS, simpleimmunostimulation may accelerate disease progression.

Immunomodulatory agents are, in general, not known for their ability totreat opportunistic infections. Further, there is presently noacceptable means for predicting whether a particular class ofimmunomodulatory agents will have utility in treating opportunisticinfections.

Badger, et al., U.S. Pat. No. 4,963,557 (Badger I) discloses compoundsof the formula I: ##STR1## wherein: n is 3-7; m is 1 or 2; R¹ and R² arethe same or different and are selected from hydrogen or straight orbranched chain alkyl, provided that the total number of carbon atomscontained by R¹ and R² when taken together is 5-10; or R¹ and R²together form a cyclic alkyl group having 3-7 carbon atoms; R³ and R⁴are the same or different and are selected from hydrogen or straightchain alkyl having 1-3 carbon atoms; or R³ and R⁴ are joined togetherwith the nitrogen atom to form a heterocyclic group having 5-8 atoms; ora pharmaceutically acceptable salt or hydrate or solvate thereof.

Badger I discloses compounds of Formula I as a class of novel compoundswhich induce an immunomodulatory effect which is characterized by thestimulation of suppressor cell activity.

Badger I does not disclose the compounds of Formula I as agents fortreating opportunistic infections.

SUMMARY OF THE INVENTION

This invention relates to a method of treating opportunistic infectionsin a mammal, including a human, in need thereof which comprisesadministering to such mammal an effective amount of a compound of theformula ##STR2## wherein: n is 3-7;

m is 1 or 2;

R¹ and R² are the same or different and are selected from hydrogen orstraight or branched chain alkyl, provided that the total number ofcarbon atoms contained by R¹ and R² when taken together is 5-10; or R¹and R² together form a cyclic alkyl group having 3-7 carbon atoms;

R³ and R⁴ are the same or different and are selected from hydrogen orstraight chain alkyl having 1-3 carbon atoms; or R³ and R⁴ are joinedtogether with the nitrogen atom to form a heterocyclic group having 5-8atoms; or a pharmaceutically acceptable salt or hydrate or solvatethereof.

DETAILED DESCRIPTION OF THE INVENTION

The preparation of all compounds of Formula (I) and pharmaceuticallyacceptable salts, hydrates and solvates and formulations thereof isdisclosed in U.S. Pat. No. 4,963,557, the entire disclosure of which ishereby incorporated by reference.

A preferred compound used in the novel method is the dihydrochloridesalt of a compound of Formula (I) where R¹ and R² are propyl, R³ and R⁴are methyl, m is 1 and n is 3 which isN,N-dimethyl-8,8-dipropyl-2-azaspiro 4.5!decane-2-propanaminedihydrochloride.

A particularly preferred compound used in the novel method is thedihydrochloride salt of a compound of Formula (I) where R¹ and R² arepropyl, R³ and R⁴ are ethyl, m is 1 and n is 3 which isN,N-diethyl-8,8-dipropyl-2-azaspiro 4.5!decane-2-propanaminedihydrochloride.

A particularly preferred compound used in the novel method is thedihydrochloride salt of a compound of Formula (I) where R¹ and R² arepropyl, R³ and R⁴ are joined together with the nitrogen to form apiperidine ring, m is 1 and n is 3 which is 8,8-dipropyl-2-azaspiro4.5!decane-2-piperidinopropyl dihydrochloride.

As used herein, the term "compound A" refers to the dihydrochloride saltof a compound of Formula (I) where R1 and R2 are propyl, R3 and R4 aremethyl, m is 1 and n is 3 which is N,N-dimethyl-8,8-dipropyl-2-azaspiro4.5!decane-2-propanamine dihydrochloride.

It has now been discovered that compounds of Formula (I) andpharmaceutically acceptable salts or hydrates or solvates thereof areuseful for treating opportunistic infections in a mammal, including ahuman, in need thereof. By the term "treating" is meant prophylactic ortherapeutic therapy.

Compound A was tested for its in vivo ability to treat opportunisticinfections in Experiment 1. To perform Experiment 1 Compound A wasdissolved in saline and administered to CBA/J mice, at 0, 1.5, or 15mg/kg, by daily intraperitoneal injection for 14 days. Control micereceived saline. On day 8, all mice received an intravenous injection of1×10⁶ Candida Albicans (Strain B311). Survival was monitored daily untilall mice died or were sacrificed for humane reasons. There was asignificant increase in the mean survival time in the mice that received15 mg/kg. In contrast, mice that received intraperitoneal 5 doses ofdexamethasone (an immunosuppressive steriod) at 50 mg/kg showed asignificant decrease in mean survival time.

Compound A was tested in an ex vivo experiment (Experiment 2) for itsability to treat opportunistic infections. In Experiment 2 Lewis ratsreceived oral doses of 20 mg/kg of Compound A dissolved in 0.5%Tragacanth (5 doses/wk) for 16 days. On day 23 the rats were sacrificedand alveolar macrophages were collected by brochoalveolar lavage. Thecells were dispensed into 24 well dishes and their ability to killCandida albicans evaluated. There was a significant increase in theability of cells from rats treated with Compound A to kill two strains(B311 and B792) of candida.

Compound A was tested in an in vitro experiment (Experiment 3) for itsability to treat opportunistic infections. In Experiment 3 alveolarmacrophges were collected by lavage from untreated rats and wereincubated with Compound A in vitro for 3 days. At that time, thecompound was washed off and the cells ability to kill Candida albicans(Strain B792) was evaluated. There was a statistically significantconcentration dependent increase in kill percentage at ≧2 uM Compound A.This effect is not due to a direct effect of Compound A on the growth ofCandida albicans as concentrations up to 12 uM had no effect on theyeast.

This invention relates to a method of treating opportunistic infectionsin a mammal, including a human, in need thereof which comprisesadministering to such mammal an effective amount of a compound ofFormula (I) or a pharmaceutically acceptable salt or hydrate or solvatethereof. A compound of Formula (I) or a pharmaceutically acceptable saltor hydrate or solvate thereof can be administered to such mammalincluding a human, in a conventional dosage form prepared by combining acompound of Formula (I) or a pharmaceutically acceptable salt or hydrateor solvate thereof, with a conventional pharmaceutically acceptablecarrier or diluent according to known techniques, such as thosedescribed in Badger (I), U.S. Pat. No. 4,963,557.

It will be recognized by one of skill in the art that the form andcharacter of the pharmaceutically acceptable carrier or diluent isdictated by the amount of active ingredient with which it is to becombined, the route of administration and other well-known variables. Acompound of Formula (I) or a pharmaceutically acceptable salt or hydrateor solvate thereof is administered to a mammal, including a human in anamount sufficient to treat opportunistic infections.

The route of administration of the Formula (I) ("active ingredient")compound is not critical but is usually oral or parenteral, preferablyoral.

The term parenteral as used herein includes intravenous, intramuscular,subcutaneous, intranasal, intrarectal, transdermal, intravaginal orintraperitoneal administration. The subcutaneous and intramuscular formsof parenteral administration are generally preferred. The dailyparenteral dosage regimen will preferably be from about 0.01 mg/kg toabout 10 mg/kg of total body weight, most preferably from about 0.1mg/kg to about 1 mg/kg. Preferably, each parenteral dosage unit willcontain the active ingredient in an amount of from about 0.1 mg to about100 mg.

The compounds of Formula (I) which are active when given orally can beformulated as liquids, for example syrups, suspensions or emulsions,tablets, capsules and lozenges.

A liquid formulation will generally consist of a suspension or solutionof the compound or pharmaceutically acceptable salt in a suitable liquidcarrier(s) for example, ethanol, glycerine, non-aqueous solvent, forexample polyethylene glycol, oils, or water with a suspending agent,preservative, flavoring or coloring agent.

A composition in the form of a tablet can be prepared using any suitablepharmaceutical carrier(s) routinely used for preparing solidformulations. Examples of such carriers include magnesium stearate,starch, lactose, sucrose and cellulose.

A composition in the form of a capsule can be prepared using routineencapsulation procedures. For example, pellets containing the activeingredient can be prepared using standard carriers and then filled intoa hard gelatin capsule; alternatively, a dispersion or suspension can beprepared using any suitable pharmaceutical carrier(s), for exampleaqueous gums, celluloses, silicates or oils and the dispersion orsuspension then filled into a soft gelatin capsule.

The daily oral dosage regimen will preferably be from about 0.01 mg/kgto about 10 mg/kg of total body weight. Preferably each oral dosage unitwill contain the active ingredient in an amount of from about 0.1 mg toabout 100 mg.

While it is possible for an active ingredient to be administered alone,it is preferable to present it as a pharmaceutical formulation.

It will be recognized by one of skill in the art that the optimalquantity and spacing of individual dosages of a compound of formula (I)or a pharmaceutically acceptable salt or hydrate or solvate thereof willbe determined by the nature and extent of the condition being treated,the form, route and site of administration, and the particular patientbeing treated, and that such optimums can be determined by conventionaltechniques. It will also be appreciated by one of skill in the art thatthe optimal course of treatment, i.e., the number of doses of a compoundof Formula (I) or a pharmaceutically acceptable salt or hydrate orsolvate thereof given per day and duration of therapy, can beascertained by those skilled in the art using conventional course oftreatment determination tests.

The method of this invention of treating opportunistic infections in amammal, including a human, comprises administering to a subject in needof such treatment an effective amount of a pharmaceutically activecompound of the present invention.

The invention also provides for the use of a compound of Formula (I) inthe manufacture of a medicament for use in the treatment ofopportunistic infections in a mammal, including a human.

The invention also provides for a pharmaceutical composition for use inthe treatment of opportunistic infections in a mammal, including ahuman.

The invention also provides for a process for preparing a pharmaceuticalcomposition containing a pharmaceutically acceptable carrier or diluentand a compound of Formula I which comprises bringing the compound ofFormula I into association with the pharmaceutically acceptable carrierof diluent.

No unacceptable toxicological effects are expected when compounds of theinvention are administered in accordance with the present invention.

Following are the results of testing the compounds of this invention.

Table I

The effect of N,N-dimethyl-8,8-dipropyl-2-azaspiro4.5!decane-2-propanamine dihydrochloride (Compound A) on treatingopportunistic infections ex vivo from Experiment 2.

                  TABLE 1                                                         ______________________________________                                        Effect of Candidacidal Activity of Rat Bronchoalveolar Cells                           STRAIN OF                                                            TREATMENT                                                                              CANDIDA     OPSONIZATION                                                                              % Kill ± S.D.                             ______________________________________                                        Control  B311        none        31 ± 12                                   Control  B311        2% rat serum                                                                              56 ± 6                                    20 mg/kg B311        none        67 ± 9*                                   20 mg/kg B311        2% rat serum                                                                              84 ± 6*                                   Control  B792        none        36 ± 4                                    Control  B792        2% rat serum                                                                              79 ± 6                                    20 mg/kg B792        none         71 ± 11*                                 20 mg/kg B792        2% rat serum                                                                              94 ± 1*                                   ______________________________________                                         *Significantly greater than control, P < 0.001                           

The data in the above table, from in vivo, ex vivo and in vitroexperiments, demonstrates the unexpected therapeutic effect of compoundsof Formula I on treating opportunistic infections.

In addition, the compounds of the present invention can beco-administered with further active ingredients, such as compounds knownto treat opportunistic infections.

Without further elaboration, it is believed that one skilled in the artcan, using the preceding description, utilize the present invention toits fullest extent. The following examples are, therefore, to beconstrued as merely illustrative and not a limitation of the scope ofthe present invention in any way.

EXAMPLE 1--CAPSULE COMPOSITION

An oral dosage form for administering Formula (I) compounds is producedby filing a standard two piece hard gelatin capsule with the ingredientsin the proportions shown in Table II, below.

                  TABLE II                                                        ______________________________________                                        INGREDIENTS               AMOUNTS                                             ______________________________________                                        N,N-diethyl-8,8-dipropyl-2-azaspiro 4,5!decane-2-                                                       25 mg                                               propanamine dihydrochloride                                                   Lactose                   55 mg                                               Talc                      16 mg                                               Magnesium stearate         4 mg                                               ______________________________________                                    

EXAMPLE 2--INJECTABLE PARENTERAL COMPOSITION

An injectable form for administering Formula (I) compounds is producedby stirring 1.5% by weight of N,N-diethyl-8,8-dipropyl-2-azaspiro4,5!decane-2-propanamine dihydrochloride in 10% by volume propyleneglycol in water.

EXAMPLE 3--TABLET COMPOSITION

The sucrose, calcium sulfate dihydrate and Formula (I) compound shown inTable III below, are mixed and granulated in the proportions shown witha 10% gelatin solution. The wet granules are screened, dried, mixed withthe starch, talc and stearic acid, screened and compressed into atablet.

                  TABLE III                                                       ______________________________________                                        Ingredients              Amounts                                              ______________________________________                                        N,N-diethyl-8,8-dipropyl-2-azaspiro 4,5!decane-                                                        20 mg                                                2-propanamine dihydrochloride                                                 calcium sulfate dihydrate                                                                              30 mg                                                sucrose                   4 mg                                                starch                    2 mg                                                talc                      1 mg                                                stearic acid             0.5 mg                                               ______________________________________                                    

While the above descriptions and examples fully describe the inventionand the preferred embodiments thereof, it is understood that theinvention is not limited to the particular disclosed embodiments comingwithin the scope of the following claims.

What is claimed is:
 1. A method for treating opportunistic infectionscaused by an organism selected from the group consisting of Candida sp.,Pseudomonas sp., Listeria sp., Pneumocystis carinii, Pneumococci,Neisseria sp., Salmonella sp., Mycobacteria sp., Cryptococcus sp.,Aspergillis sp., Cryptosporidium sp., Herpes simplex, Herpes zoster,Cytomegalovirus and Toxoplasma sp. in a mammal including a human subjectto such infection which comprises administering to said human aneffective amount of a compound of the formula ##STR3## wherein: n is3-7;m is 1 or 2; R¹ and R² are the same or different and are selectedfrom hydrogen or straight or branched chain alkyl, provided that thetotal number of carbon atoms contained by R¹ and R² when taken togetheris 5-10; or R¹ and R² together form a cyclic alkyl group having 3-7carbon atoms; R³ and R⁴ are the same or different and are selected fromhydrogen or straight chain alkyl having 1-3 carbon atoms; or R³ and R⁴are joined together to form a cyclic alkyl group having 4-7 atoms; or apharmaceutically acceptable salt or hydrate or solvate thereof.
 2. Themethod of claim 1 wherein the compound isN,N-diethyl-8,8-dipropyl-2-azaspiro 4.5!decane-2-propanamine; or apharmaceutically acceptable salt, hydrate or solvate thereof.
 3. Themethod of claim 1 wherein the compound is administered orally.
 4. Themethod of claim 3 wherein from about 0.01 mg/kg to about 10 mg/kg ofcompound is administered per day.
 5. The method of claim 1 wherein thecompound is administered parenterally.
 6. The method of claim 5 whereinfrom about 0.01 mg/kg to about 10 mg/kg of compound is administered perday.